薄荷茉莉酸信号抑制子McJAZ8基因的鉴定与功能分析

Identification and functional analysis of jasmonic acid signaling repressor McJAZ8 gene in Mentha canadensis

薄荷作为药食同源植物,具有重要经济价值。茉莉酸信号抑制子JAZ蛋白在调节植物应答逆境胁迫方面具有重要作用。克隆薄荷McJAZ8基因,并分析其蛋白特征、蛋白互作和表达模式,为薄荷抗逆分子育种提供基因资源。该实验将通过生物信息学、酵母双杂交、烟草叶片瞬时表达、qRT-PCR等技术对McJAZ8的蛋白结构特征、亚细胞定位、蛋白互作与基因表达进行分析。结果显示:①McJAZ8基因全长543 bp,编码180个氨基酸,McJAZ8蛋白具有保守的TIFY和Jas结构域,与拟南芥AtJAZ1和AtJAZ2同源性较高。②McJAZ8蛋白定位于细胞核和细胞质。③酵母双杂交结果显示,McJAZ8与其自身或McJAZ1/3/4/5蛋白互作形成同源或异源二聚体。④McJAZ8在不同组织中均有表达,幼叶中表达量最高。在叶序中,McJAZ8在第4片叶中表达量最高,而第2片叶中表达量最低。⑤在叶片和根中,McJAZ8在茉莉酸甲酯(MeJA)、干旱、NaCl处理下的表达呈不同程度地上调;McJAZ8的表达在CdCl2处理下呈先上调而后下调的模式;在叶片中,McJAZ8的表达在CuCl2处理下呈逐渐下调的趋势,而在根中呈先下调后上调再下调的趋势;在叶片和根中,McJAZ8在AlCl3处理下的表达均呈不同程度的下调。该研究丰富了薄荷中茉莉酸信号抑制子JAZ基因的研究,并为薄荷分子育种提供了基因资源。

Mentha canadensis,as a plant with medicinal and culinary uses,holds significant economic value.Jasmonic acid signaling repressor JAZ protein has a crucial role in regulating plant response to adversity stresses.The M.canadensis McJAZ8 gene is cloned and analyzed for protein characterization,protein interactions,and expression patterns,so as to provide genetic resources for molecular breeding of M.canadensis for stress tolerance.This experiment will analyze the protein structural characteristics,subcellular localization,protein interactions,and gene expression of McJAZ8 using bioinformatics,yeast two-hybrid(Y2H),transient expression in tobacco leaves,qRT-PCR,and other technologies.The results show that:①The full length of the McJAZ8 gene is 543 bp,encoding 180 amino acids.The McJAZ8 protein contains conserved TIFY and Jas domains and exhibits high homology with Arabidopsis thaliana AtJAZ1 and AtJAZ2.②The McJAZ8 protein is localized in the nucleus and cytoplasm.③The Y2H results show that McJAZ8 interacts with itself or McJAZ1/3/4/5 proteins to form homologous or heterologous dimers.④McJAZ8 is expressed in different tissue,with the highest expression level in young leaves.In terms of leaf sequence,McJAZ8 shows the highest expression level in the fourth leaf and the lowest expression level in the second leaf.⑤In leaves and roots,the expression of McJAZ8 is upregulated to varying degrees under methyl jasmonate(MeJA),drought,and NaCl treatments.The expression of McJAZ8 shows an initial upregulation followed by a downregulation pattern under CdCl2 treatment.In leaves,the expression of McJAZ8 tends to gradually decrease under CuCl2 treatment,while in roots,it initially decreases and then increases before decreasing again.In both leaves and roots,the expression of McJAZ8 is downregulated to varying degrees under AlCl3 treatment.This study has enriched the research on jasmonic acid signaling repressor JAZ genes in M.canadensis and provided genetic resources for the molecular breeding of M.canadensis.